Vibrio vulnificus is an important foodborne pathogenic bacterium that mainly contaminates seafood. Rapid and accurate technologies that suitable for on-site detection are critical for effective control of its spreading. Conventional detection methods and polymerase chain reaction (PCR)-based and qPCR-based approaches have application limitations in on-site scenarios. Application of loop-mediated isothermal amplification (LAMP) technology was a good step towards the on-site detection. In this study, a recombinase polymerase amplification (RPA)-based detection method for V. vulnificus was developed combining with lateral flow strip (LFS) for visualized signal. The method targeted the conservative empV gene encoding the extracellular metalloproteinase, and finished detection in 35 min at a conveniently low temperature of 37 °C. It showed good specificity and an excellent sensitivity of 2 copies of the genome or 10⁻¹ colony forming unit (CFU) per reaction, or 1 CFU/10 g in spiked food samples with enrichment. The method tolerated unpurified templates directly from sample boiling, which added the convenience of the overall procedure. Application of the RPA-LFS method for clinical samples showed accurate and consistent detection results compared to bioassay and quantitative PCR. This RPA-LFS combined method is well suited for on-site detection of V. vulnificus.

创伤弧菌是一种重要的食源性致病细菌，主要污染海鲜。适用于现场检测的快速而准确的技术对于有效控制其传播至关重要。常规检测方法以及基于聚合酶链反应（PCR）和基于qPCR的方法在现场场景中具有应用限制。回路介导的等温扩增（LAMP）技术的应用是朝现场检测迈出的良好一步。在这项研究中，开发了一种基于重组酶聚合酶扩增（RPA）的创伤弧菌检测方法，并结合了横向流条（LFS）来显示可视化信号。该方法针对保守的empV基因编码细胞外金属蛋白酶，并在方便的37°C低温下于35分钟内完成检测。它显示出良好的特异性和极好的灵敏度，每个反应2个基因组拷贝或10 ^-1个菌落形成单位（CFU），或加标食品样品中1 CFU / 10 g。该方法可直接从样品沸腾中耐受未纯化的模板，从而增加了整个过程的便利性。与生物测定和定量PCR相比，RPA-LFS方法在临床样品中的应用显示出准确一致的检测结果。这种RPA-LFS组合方法非常适合于现场检测V. vulnificus。